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Organization and Evolution of 5S Ribosomal Dna in the Fish Genome

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Organization and Evolution of 5S Ribosomal Dna in the Fish Genome In: Focus on Genome Research ISSN 1-59033-960-6 Editor: Clyde R. Williams, pp335-363 ©2004 Nova Science Publishers, Inc. Chapter X ORGANIZATION AND EVOLUTION OF 5S RIBOSOMAL DNA IN THE FISH GENOME Cesar Martins 1 and Adriane Pinto Wasko 2 Departamento de Morfologia, Instituto de Biociências, Universidade Estadual Paulista, CEP 18618-000, Botucatu, SP, Brazil. Phone/Fax +55 14 38116264, 1e-mail [email protected]; 2e-mail: [email protected] ABSTRACT In higher eukaryotes, the 5S ribosomal multigene family (5S rDNA) is tandemly organized in repeat units composed of a coding region (5S rRNA gene) and a non-transcribed spacer sequence (NTS). Although the 5S rDNA organization has been investigated in several vertebrate species, present data are concentrated in mammals and amphibians, whereas other groups, such as fishes, have been poorly studied. To further the understanding on the dynamics and evolution of 5S rDNA arrays in the vertebrate genome, recent studies have focused on the genome organization of these sequences in fish species, which represent the base group of vertebrate evolution. It was evidenced that the chromosome distribution of the 5S rDNA is quite conserved among related fish species occupying an interstitial position in the chromosomes. Although the 5S rDNA clusters have been maintained conserved in the chromosomes, changes in the nucleotide sequences and organization of the repeat units have occurred in fish species, as demonstrated by the presence of 5S rDNA variant types within and between genomes clustered in distinct chromosome environments. These variants are distributed in two major classes, suggesting that such pattern could represent a primitive condition for the fish genome, as well as for vertebrates. The 5S rDNA variant units include the presence of pseudogenes and inverted gene sequences, demonstrating that the organization of the 5S rDNA repeats in fish genome is governed by intense mechanisms of evolution. Data on the 5S rDNA repeat organization can provide interesting insights for the comprehension of (1) the genome evolution, (2) the dynamics of repetitive sequences, and (3) the practical employment of 5S rDNA as genetic markers. 336 C. Martins and A. P. Wasko INTRODUCTION In general, little is known about the structure and organization of fish genome - most available information in this vertebrate class is related to the structure and evolution of chromosomes. Molecular studies focusing on genes and DNA sequences of this animal group are mainly related to repetitive sequences such as satellite and ribosomal DNAs. Single copy sequences correspond to approximately 2-10% of the vertebrate genomes, while the remainder genome is composed by repetitive sequences (Franck et al. 1991, Wagner et al. 1993, Bernardi 1995). Repetitive DNA includes the tandemly-arrayed satellite, minisatellite and microsatellite sequences, and dispersed repeats such as transposons and retrotransposons (Charlesworth et al. 1994). There are also repetitive multigene families that code several important proteins such as hemoglobins, actins, histones, and ribosomal RNAs (rRNA). While the non-coding tandemly-arrayed sequences are found mainly in chromosome centromeres and telomeres, the dispersed elements are found as isolated units or as clusters distributed throughout the genome. The rRNA genes can be found clustered in one or several regions of the genome. The ribosomal RNA is the most abundant RNA in the cell, constituting approximately 80% of the total RNA of diving cells. rRNAs represent noncoding final gene products that perform structural and catalytic functions and form the basic structure of the large and small ribosomal subunits that catalyze cellular protein synthesis. In animals, the large ribosome subunit is formed by the association of one copy of each rRNA (28S, 5.8S and 5S) with around 49 ribosomal proteins, and the small subunit is formed by the association of one 18S rRNA copy with around 33 ribosomal proteins. The synthesis of a specific protein requires multiple translation rounds of its messenger RNA molecule providing a large amount of such protein in the cell. For such purpose, adequate quantities of the ribosomal subunits are necessary and high levels of rRNA molecules are required. Large quantities of ribosomal molecules are produced just because the cell contains multiple copies of the rRNA genes that are arranged into several genomic locations. In higher eukaryotes, tandem arrays of ribosomal RNA genes are organized in multigene families. Multigene families correspond to a common structural element of the eukaryote genomes and are defined as a set of genes derived by duplication of an ancestral gene and that display a similarity of more than 50%. They are frequently found in close linkage with each other, and have similar or overlapping functions. The head-to-tail array of the rRNA multigene families tandem repeats confers a particular dynamic to the evolution of these genome regions, making them important for the comprehension of the genome structure and evolution. Studies on ribosomal RNA genes have been gaining prominence in a broad range of animal and plants, specially related to species or population characterization, evolutionary relationships and gene expression. In higher eukaryotes, the tandem arrays of rRNA genes are organized in two distinct multigene families (Figure 1) composed of hundreds to thousands of copies. These rRNA molecules are codified by two multigene families that are organized in two distinct repeat classes denominated 45S ribosomal DNA (rDNA) and 5S rDNA. The 45S rDNA contains the genes that code for the 18S, 5.8S and 26S-28S rRNAs (see Long and David 1980). The second ribosomal family codifies the 5S rRNA. The 18S, 5.8S and 26S-28S rRNAs are synthesized, processed and partially assembled to form the ribosome subunits in Organization And Evolution of 5S Ribosomal DNA In The Fish Genome 337 the nucleolus. On the other hand, the 5S rRNA is synthesized elsewhere in the genome and enters the nucleolus to participate in the large ribosomal subunit assemblage. The general nucleotide sequence and mapping pattern of these two multigene families have contributed to the comprehension of the structure, organization and evolution of the genomes. plants 18S 5,8S 26S 5’ ITS 1 ITS 2 IGS 5S 5’ NTS animals 18S 5,8S 28S 5’ ITS 1 ITS 2 IGS 5S 5’ NTS fungi and protists 18S 5,8S 28S 5S 5’ ITS 1 ITS 2 fungi, nematodes, and crustaceans 18S 5’ ITS 1 ITS 2 5S protists and nematodes 5’ TSL 5S 5’ TSL 5S crustacean histone genes 5’ 5S fungi and protists 5S 5S 5S 5S 5S 5S Figure 1. Graphical representation of the 5S rRNA genes organization and their relation to the 18S, 5,8S and 28S rRNA genes and other repeat units. The boxes represent the coding region of different genes. Boxes above the line demonstrate that the 5S rRNA gene is transcribed from the opposite strand of the double DNA helix. The 5S rRNA genes are generally organized in a distinct locus from the other rRNA genes as observed for most plant and animals, or dispersed in the genome as observed in fungi and protists. On the other hand, 5S rRNA genes can also be linked to the 18S, 5,8S and 28S rRNAs (protists, fungi, nematodes, and crustaceans), TSL (protists and nematodes) or histone genes (crustacean). ITS, internal transcribed spacer; IGS, intergenic spacer; NTS, non-transcribed spacer; TSL, trans-spliced leader. A considerable amount of information on the structural and functional organization of the 5S rRNA genes has been gained for plants (Hanson et al. 1996, Adacchi et al. 1997, Amarasinghe and Carlson 1998, among others), mammals (Little and Braaten 1989, Leah et al. 1990, Suzuki et al. 1994) and some amphibian species (Korn 1982, Vitelli et al. 1982, del Pino et al. 1992). In most eukaryotes, the 5S rRNA genes are generally detected in distinct areas of the genome, organized as one or more tandemly repeated clusters. The number of 5S rRNA gene copies ranges from 100 to 300,000, which is usually higher than the number of the 45S rRNA genes (Hadjiolov 1985). While in Drosophila , for instance, there is only one cluster of about 160 5S rRNA gene copies, in Xenopus laevis the 5S rRNA gene clusters are 338 C. Martins and A. P. Wasko located at the telomeric region of most, if not all, chromosomes (Pardue et al. 1973). On the other hand, in the fungi Neurospora crassa (Selker et al. 1981) and Schizosaccharomyces pombe (Mao et al. 1982) and in the protist Acanthamoeba castellanii (Zwick et al. 1991) the 5S rRNA gene copies are dispersed all over the genome. The evolution seems to have lead to a clustered arrangement of the 5S rRNA genes in higher eukaryotes. The organization and expression of the nuclear rRNA genes have been extensively studied for several organisms from bacteria to animals. The schemes of Figure 1 summarize the organization of the rRNA genes in the genome of eukaryotes. In eubacteria the rRNA genes coding for 16S, 23S, and 5S rRNA are usually organized in operons and are transcribed in this order by a single RNA polymerase. In contrast, eukaryotes have four different rRNA genes that are transcribed by two distinct RNA polymerases. The 18S, 5.8S, and 28S rRNA eukaryote genes are arrayed in such order to form RNA transcript units for the RNA polymerase I, and multiple copies of this units are found clustered in long direct tandem repeats. The 5S rRNA genes are transcribed by the RNA polymerase III and are also found in direct tandem repeats. The eukaryote 18S, 28S, and 5S rRNA genes are homologous to the 16S, 23S, and 5S rRNA genes of prokaryotes, respectively, whereas the 5.8S rRNA eukaryote gene is homologous to the 5’ end of the eubacterial 23S rRNA gene (Gerbi 1985) (see Figure 2). higher eukaryotes 18S 5,8S 26S/28S 5S 16S 23S 5S prokaryotes Figure 2: Homology of rRNA genes in prokaryotes and eukaryotes.
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